Taxonomic characterization of Sphaerotilus microaerophilus sp. nov., a sheath-forming microaerophilic bacterium of activated sludge origin.

A microaerophilic Gram-stain-negative bacilliform bacterial strain, FB-5 T, was isolated from activated sludge in Yokohama, Japan, that exhibited filamentous growth and formed a microtube (sheath). Cells were motile using a single polar flagellum. The optimum growth temperature and pH were 30 °C and 7.5, respectively. Strain FB-5 T was catalase-negative. Peptides and amino acids were utilized as energy and carbon sources. Sugars and organic acids were not utilized. Vitamin B12 enhanced the growth of strain FB-5 T. Sulfur-dependent lithotrophic growth was possible. Major respiratory quinone was UQ-8. Major fatty acids were C16:1ω7 and C16:0. The genomic DNA G + C content was 69.16%. Phylogenetic analysis of the 16S rRNA gene suggested that strain FB-5 T belongs to the genus Sphaerotilus. The close relatives were S. natans subsup. sulfidivorans and S. natans subsup. natans with 98.0% and 97.8% similarity based on the 16S rRNA gene analysis, respectively. The genome size (6.06 Mbp) was larger than that (4.39-5.07 Mbp) of the Sphaerotilus strains. The AAI values against the related strains ranged from 71.0 to 72.5%. The range of ANI values was 81.7 - 82.5%. In addition to these distinguishable features of the genome, the core genome and dDDH analyses suggested that this strain is a novel member of the genus Sphaerotilus. Based on its physiological properties and genomic features, strain FB-5 T is considered as a novel species of the genus Sphaerotilus, for which the name S. microaerophilus sp. nov. is proposed. The type strain is FB-5 T (= JCM 35424 T = KACC 23146 T).

Transcatheter arterial embolization for hemothorax caused by spinal fracture without arterial injury: a case report and review of the literature.

BACKGROUND: Spinal fractures rarely cause hemothorax, and no treatment consensus has been reached. Conservative treatment is generally selected in cases without arterial injury, but there have been some reports of uncontrolled bleeding. Here we report a case of hemothorax caused by spinal fracture without arterial injury treated with transcatheter arterial embolization. CASE PRESENTATION: An 88-year-old Japanese woman with back pain was diagnosed with hemothorax due to bleeding from an unstable fracture of the tenth thoracic vertebra. Contrast-enhanced computed tomography revealed no obvious arterial injury. We performed transcatheter arterial embolization of the bilateral tenth intercostal arteries to prevent rebleeding. The hemothorax did not worsen until surgical spinal fixation 9 days post-transcatheter arterial embolization, and she was discharged 30 days after admission. CONCLUSION: Transcatheter arterial embolization for hemothorax caused by spinal fractures without obvious arterial injury may be a useful bridge to spinal fixation.

Novel approach for rapid detection of extended spectrum ß-lactamase and metalloid-ß-lactamase using drug susceptibility testing microfluidic device (DSTM).

BACKGROUND/PURPOSE: Rapid detection of ß-lactamases is important in a recent situation where resistant bacteria are increasing. By using the drug susceptibility testing microfluidic device (DSTM), rapid screening of extended spectrum ß-lactamases (ESBLs) and metallo-ß-lactamases (MBLs) has become possible. METHODS: ß-lactams and ß-lactamase inhibitors were pre-fixed in the DSTM for use. A bacterial suspension in Mueller-Hinton broth (McF 0.25) was introduced into the device, and the effects of ß-lactamase inhibitor on morphological changes caused by ß-lactam were evaluated after 3 h incubation. RESULTS: Clinical isolates genetically confirmed to produce ß-lactamase were used. Of the 84 ESBL-producing strains, 80 strains (95%) turned to be ESBL positive, and five strains (6%) of them MBL were positive as well as ESBL. Four strains (5%) were negative for both ESBL and MBL. Of the 24 MBL-producing strains, 23 strains (96%) were positive for MBL. All the 43 AmpC-producing strains were negative for both ESBL and MBL. Of the 156 ESBL- and MBL-nonproducing strains, 155 strains (99%) were negative for both ESBL and MBL, and one strain was positive for ESBL. With this method, the detection sensitivity was 95% and the specificity was 100% for ESBL, whereas the detection sensitivity was 96% and the specificity was 98% for MBL. These results were not significantly different from the results of the disc diffusion method. CONCLUSION: The DSTM method allows rapid detection of ß-lactamases in 3 h and may be a useful replacement for the disc diffusion method.

Extremely high troponin levels induced by septic shock: a case report.

BACKGROUND: Troponin levels can be elevated in various diseases other than acute myocardial infarction, including sepsis. In diseases without myocardial necrosis, the elevated troponin levels are relatively low and normalize quickly. CASE PRESENTATION: A 61-year-old Japanese man with impaired consciousness was transported to our hospital. He was diagnosed as having pneumonia and septic shock. His condition was severe, but his clinical course was good. However, his troponin level remained extremely high during admission; on the second day, it was higher than the measurable range. We consulted a cardiologist and performed echocardiography and myocardial perfusion scintigraphy but found no new ischemic changes. CONCLUSION: In septic shock, troponin levels can be extremely high, which can persist even after recovery, as in very large myocardial infarctions.

Ultraselective transcatheter arterial embolization with small-sized microcoils for acute lower gastrointestinal bleeding.

PURPOSE: To evaluate the clinical outcome of ultraselective transcatheter arterial embolization (TAE) with small-sized microcoils for acute lower gastrointestinal bleeding (LGIB). MATERIALS AND METHODS: The subjects were 17 consecutive patients (mean age, 69 years) with LGIB who were treated with ultraselective TAE using small-sized microcoils between December 2013 and December 2019. Ultraselective TAE was defined as embolization of one or both of the long or short branches of the vasa recta. The etiologies of bleeding were colonic diverticulosis in 16 patients (94%) and malignancy in one patient (6%). The bleeding foci were in the ascending colon in 11 patients (65%), transverse colon in 2 patients (12%), and sigmoid colon in 4 patients (23%). A total of 18 branches (diameter: range 0.5-1.5 mm, mean 1.1 mm) of the vasa recta in 17 patients were embolized with small-sized microcoils (size range 1-3 mm, mean combined lengths of all microcoils 7.6 cm). The mean follow-up period was 19 months (range 1-80 months). The technical and clinical success rate, recurrent bleeding rate, major complications and long-term clinical outcomes were retrospectively evaluated. RESULTS: Technical and clinical success was achieved in all patients (17/17). The rates of early recurrent bleeding (within 30 days of TAE) and major complications were 0% (0/17). Recurrent bleeding occurred in one patient at 2 months after TAE, but was stopped with conservative treatment. There were no other bleeding episodes or complications in the follow-up period. CONCLUSION: Ultraselective TAE with small-sized microcoils is a highly effective and safe treatment modality for LGIB.

A physician-staffed ground emergency medical service does not significantly shorten door-to-balloon time in patients with STEMI: an observational study in a single emergency center in Japan.

AIM: Current guidelines recommend a door-to-balloon time (DTBT) of <90 min for reperfusion treatment of patients with ST-segment elevation myocardial infarction (STEMI). A physician-staffed ground emergency medical service (GEMS) using a rapid response car (RRC) system was implemented at our hospital in April 2015. The medical team, including a physician and nurse, is dispatched to assess the patient and expedite the start of treatment by emergency physicians and cardiologists after arrival at the hospital. The study aimed to determine whether the RRC system shortened the DTBT. METHODS: This retrospective observational study was carried out in a tertiary emergency center in Japan. Those STEMI patients with primary percutaneous intervention between January 2016 and December 2018 were evaluated. The DTBTs of patients transported by the RRC system, the emergency medical service (EMS), and transferred from other hospitals after STEMI diagnosis (TRANS group) were compared. RESULTS: A total of 121 patients were included, 33 in the RCC, 20 in the EMS, and 68 in the TRANS groups. The median DTBT was 51 min (interquartile range [IQR], 43-67) in the RRC, 61 min (IQR, 52-85) in the EMS, and 59 min (IQR, 48-72) in the TRANS groups (P = 0.13). The DTBT was not significantly shorter in the RRC than in the other groups. CONCLUSION: An RRC physician-staffed GEMS did not significantly shorten the DTBT of patients with STEMI compared with other transport systems.

A rapid response-type doctor car system shortened time to intravenous thrombolytic therapy for patients with ischemic stroke: an observational study at a single emergency center in Japan.

BACKGROUND: For patients with ischemic stroke, rapid reperfusion therapy is extremely important. In April 2015, our medical center introduced a rapid response-type doctor car (RRC) system. Here, an emergency medical team, including a physician, is dispatched to the patient's prehospital location. The team then assesses the patient and, if necessary, initiates infusion therapy, excluding thrombolytic therapy. Before arriving at the hospital, a prehospital physician orders the preparation of diagnostic tools and conducts an early consultation to a neurologist in order to begin thrombolytic therapy more swiftly. This study aimed to determine whether the RRC system shortened the time to commence intravenous reperfusion therapy in patients with ischemic stroke. This was a retrospective observational study conducted at a tertiary emergency center in Japan. Cases of patients with ischemic stroke who underwent intravenous thrombolytic therapy from January 2015 to December 2018 were enrolled. They were divided into two groups: RRC group (intervened by RRC system) and non-RRC group (not intervened by RRC system). The groups' door-to-needle (DTN) time was compared. RESULTS: During the study period, 140 patients received intravenous thrombolytic therapy. Among those, 28 were in the RRC group and 28 received the usual prehospital care. Of 56 patients, the median age was 82 years old, and 42.9% of patients were male. The median NIHSS was 14 (IQR 10-21). As for demographics, there were no significant differences between the two groups. Median DTN time was 67 min (IQR 55-79) in RRC group vs. 81 min (IQR 69-107) in usual care group, respectively (P < 0.05). CONCLUSION: In this study, patients with ischemic stroke in RRC group received intravenous thrombolytic therapy in a shorter time compared to the group that received usual care.

Heterobimetallic triple-decker complexes derived from a dianionic aromatic stannole ligand.

A neutral heterobimetallic triple-decker stannole complex was prepared by the reaction of an anionic ruthenocene bearing a stannole dianionic ligand with [Rh(cod)Cl]2 (cod = 1,5-cyclooctadiene), and the resulting Ru-Rh complex exhibits an electronic property different from those of the corresponding Ru-Ru and Rh-Rh complexes. The Ru-Rh complex can be decomposed in ionic liquids to metal nanoparticles.

Discovery of a Slow Tight Binding LPA1 Antagonist (ONO-0300302) for the Treatment of Benign Prostatic Hyperplasia.

Scaffold hopping from the amide group of lead compound ONO-7300243 (1) to a secondary alcohol successfully gave a novel chemotype lysophosphatidic acid receptor 1 (LPA1) antagonist 4. Wash-out experiments using rat isolated urethra showed that compound 4 possesses a tight binding feature to the LPA1 receptor. Further modification of two phenyl groups of 1 to pyrrole and an indane moiety afforded an optimized compound ONO-0300302 (19). Despite its high i.v. clearance, 19 inhibited significantly an LPA-induced increase of intraurethral pressure (IUP) in rat (3 mg/kg, p.o.) and dog (1 mg/kg, p.o.) over 12 h. Binding experiments with [3H]-ONO-0300302 suggest that the observed long duration action is because of the slow tight binding character of 19.

Discovery of a novel EP2/EP4 dual agonist with high subtype-selectivity.

A series of γ-lactam prostaglandin E(1) analogs bearing a 16-phenyl moiety in the ω-chain and aryl moiety in the α-chain were synthesized and biologically evaluated. Among the tested compounds, γ-lactam PGE analog 3 designed as a structural hybrid of 1 and 2 was discovered as the most optimized EP2/EP4 dual agonist with excellent subtype-selectivity (K(i) values: mEP2=9.3 nM, mEP4=0.41 nM). A structure-activity relationship study is presented.

Adsorption of lecithin liposomes to acid clay.

The interaction between lecithin liposomes and acid clay was investigated to clarify the mechanism for liposome adsorption to the clay. It was found that the multilamellar vesicular structure of the liposomes was broken as a result of primary adsorption. The acid clay particles aggregated and were eventually covered by the lecithin layer structure. In the case of kaolin, on the other hand, the liposomes were weakly adsorbed to the clay and maintained the vesicular structure. The amount of primary adsorption to the clay surface, which was estimated from the adsorption isotherm, was more for acid clay than for kaolin, and the total amount adsorbed to the acid clay was also more than to kaolin. This result can be explained by the much higher density of the negative charge on the acid clay surface than that for kaolin. The liposomes are therefore considered to be adsorbed to the acid clay mainly by the choline positive charge residing at the end of the lecithin molecule, although this is of no net charge as a whole.

Spirodiketopiperazine-based CCR5 antagonist: discovery of an antiretroviral drug candidate.

Following the discovery that hydroxylated derivative 3 (Fig. 1) was one of the oxidative metabolites of the original lead 1, it was found that hydroxylated compound 4 possesses higher in vitro anti-HIV potency than the corresponding non-hydroxylated compound 2. Structural hybridation of 4 with the orally available analog 5 resulted in another orally-available spirodiketopiperazine CCR5 antagonist 6a that possesses more favorable pharmaceutical profile for use as a drug candidate.

Culture scale-up studies as seen from the viewpoint of oxygen supply and dissolved carbon dioxide stripping.

Oxygen supply and dissolved carbon dioxide (dCO(2)) stripping are two of the most important control parameters in cell culture. In this study, we investigated the effect of scale-up on the volumetric gas transfer coefficient with bioreactors of different sizes (working volume: 80 L, 500 L, 2000 L, and 10,000 L; aspect ratio: 1.0-1.6). Sparging air into water increased the volumetric oxygen transfer coefficient (k(L)a), an index of oxygen supply efficiency, by scale-up roughly in proportion to the depth of the water. A corresponding increase in k(L)a was found in a real cell culture of Chinese hamster ovary cells. dCO(2) stripping efficiency was evaluated in water tests using changes in k(L)a(co2), an index defined in relation to k(L)a. k(L)a(co2) increased following surface aeration, but the rate of increase was reduced by scale-up, which was attributed to a decrease in the liquid surface-to-volume ratio. A similar decrease in efficiency was observed in a 2000 L bioreactor by increasing the liquid volume at constant liquid surface area. The observed scale-up effects are discussed based on a simple theoretical consideration.

Estimation of dissolved carbon dioxide stripping in a large bioreactor using model medium.

Dissolved carbon dioxide (dCO(2)) accumulation is one of the most serious problems in the scale-up of industrial cell culture. To predict the effects of dCO(2) stripping in different culture conditions and at different scales, we examined a method of estimation of dCO(2) stripping using a model medium. The operational parameters (e.g., sparging and agitation rate) and the size of the bioreactor (working volume: 80 L, 500 L, 2000 L; aspect ratio: 1.0 approximately 1.6) were varied, and the model medium was prepared by adjusting pH, density, viscosity, surface tension, and buffer conditions. dCO(2) stripping efficiency was evaluated using the index k(L)a(CO)((2)), which was defined in accordance with the volumetric oxygen transfer coefficient k(L)a. The model medium exhibited dCO(2) stripping behavior similar to real culture medium in all experimental conditions tested. It is expected that the use of the model medium to estimate dCO(2) stripping in real cultures will be valuable for determining the culture conditions in bioreactors in scale-up.

Spirodiketopiperazine-based CCR5 antagonists: Lead optimization from biologically active metabolite.

Hydroxylated derivatives were designed and synthesized based on the information of oxidative metabolites. Compounds derived from beta-substituted (2R,3R)-2-amino-3-hydroxypropionic acid showed improved inhibitory activities against the binding of MIP-1alpha to human CCR5, compared with the non-hydroxylated derivatives and the other isomers.

Improvement by solid dispersion of the bioavailability of KRN633, a selective inhibitor of VEGF receptor-2 tyrosine kinase, and identification of its potential therapeutic window.

KRN633 is a potent inhibitor of vascular endothelial growth factor (VEGF) receptor tyrosine kinases. However, it is poorly water-soluble; consequently, relatively high doses are required to achieve substantial in vivo tumor growth suppression after oral administration. We subjected KRN633 to the solid dispersion technique to improve its solubility, absorption, and antitumor efficacy after oral administration. This technique transformed the drug into an amorphous state and dramatically improved its dissolution rate. It also enhanced the bioavailability of the drug in rats by approximately 7.5-fold. The solid dispersion form of KRN633 also dramatically inhibited human tumor growth in murine and rat xenograft models: similar rates of tumor growth inhibition were obtained with 10- to 25-fold lower doses of the solid dispersion preparation relative to the pure drug in its crystalline state. Histologic analysis of tumors treated with the solid dispersion preparation revealed a significant reduction in microvessel density at much lower doses when compared with the crystalline form preparation. In addition, a dose-finding study using the solid dispersion form in a rat xenograft model revealed that there was a substantial range of doses at which KRN633 in the solid dispersion form showed significant antitumor activity but did not induce weight loss or elevate total urinary protein levels. These data suggest that the solid dispersion technique is an effective approach for developing KRN633 drug products and that KRN633 in the solid dispersion form may be a highly potent, orally available drug with a wide therapeutic window for diseases associated with abnormal angiogenesis.

Discovery of novel phosphonic acid derivatives as new chemical leads for inhibitors of TNF-alpha production.

2-(Acylamino)benzylphosphonic acid 6 derived from an artificial substrate of sphingomyelinase was found to show inhibitory activity of TNF-alpha production. Structural optimization was started with the chemical modification of 6. The discovery of another chemical leads 7, 8, 10 and 16 for the development of structurally new inhibitors of TNF-alpha production is reported.

Melatonin alleviates jet lag symptoms caused by an 11-hour eastward flight.

The effect of 3 mg of melatonin on the rate of re-entrainment of plasma melatonin rhythm after an 11-h eastward flight was assessed. Eight subjects participated in the study, and underwent 24-h blood samplings once before the flight and twice after the flight. Subjects were exposed to natural zeitgeber outdoors and took 3 mg of malatonin at 20:00 h local time on the days when no blood sampling was done. Antidromic re-entrainment was dominant whereby melatonin administration in the evening promoted re-entrainment. Melatonin accelerated the rate of re-entrainment by 15 min per day and alleviated the jet lag symptoms.